Rotatable sample disk and method of loading a sample disk

ABSTRACT

A rotatable sample disk configured for samples of biological material. The sample disk may include a fill chamber for storing a first biological material, a plurality of first sample chambers positioned in the sample disk farther from the rotational axis of the sample disk than the fill chamber, a plurality of second sample chambers, and a plurality of circumferential fill channels. Each of the second sample chambers may be configured to permit fluid communication with a respective first sample chamber. The plurality of circumferential fill conduits may be configured to permit transfer of the first biological material from the fill chamber to the plurality of first sample chambers upon a first rotation of the sample disk about the rotational axis. Methods of loading a plurality of sample chambers in a sample disk are also provided.

FIELD

[0001] The present teachings relate generally to a sample diskconfigured for samples of biological material, and methods of loading asample disk. The present teachings further relate, in various aspects,to a sample disk that is rotatable about a rotational axis in order tocentrifugally load sample chambers of the sample disk with biologicalmaterial.

BACKGROUND

[0002] Biological testing has become an important tool in detecting andmonitoring diseases. In the biological testing field, thermal cycling isused to amplify nucleic acids by, for example, performing polymerasechain reactions (PCR) and other reactions. PCR, for example, has becomea valuable research tool with applications such as cloning, analysis ofgenetic expression, DNA sequencing, and drug discovery. Methods such asPCR may be used to detect a reaction of a test sample to ananalyte-specific reagent. Typically, an analyte-specific reagent isplaced in each sample chamber in advance of performing the testing. Thetest sample is then later inserted into the sample chambers, and thesample well tray or microcard is then transported to a thermal cyclingdevice.

[0003] Recent developments in the field have led to an increased demandfor biological testing devices. Biological testing devices are now beingused in an increasing number of ways. It is desirable to provide a moreefficient and compact method and structure for filling and thermallycycling substrates such as sample trays and microcards.

[0004] In typical systems, the sample tray or microcard is loaded withreagent, then loaded with the test sample, and then transported andinserted into a separate device for thermal cycling. It is desirable toreduce the amount of time and number of steps taken to fill andthermally cycle a sample tray or microcard.

SUMMARY

[0005] Various aspects generally relate to, among other things, arotatable sample disk configured for samples of biological material.According to one various aspects, the sample disk can include a fillchamber for storing a first biological material, a plurality of firstsample chambers positioned in the sample disk farther from a rotationalaxis than the fill chamber, a plurality of second sample chambers, and aplurality of circumferential fill conduits positioned adjacent theplurality of first sample chambers. In various embodiments, the fillchamber is configured for rotation on the sample disk about a rotationalaxis. Each of the second sample chambers may be configured to permitfluid communication with a respective first sample chamber. The secondsample chambers may be positioned closer to the rotational axis than thefirst sample chambers. The plurality of circumferential fill channelsmay be configured to permit transfer of the first biological materialfrom the fill chamber to the plurality of first sample chambers upon afirst rotation of the sample disk about the rotational axis.

[0006] Various aspects comprise a method of loading a plurality ofsample chambers on a sample disk. The method can include the step ofproviding a sample disk with a fill chamber, a plurality of first samplechambers, and a plurality of second sample chambers. The method mayfurther comprise loading the plurality of first sample chambers with afirst biological material by rotating the sample disk about a rotationalaxis so that a first biological material in the fill chamber travelsthrough a plurality of circumferential fill conduits connecting the fillchamber with the first sample chambers. The plurality of circumferentialfill conduits may be positioned between adjacent first sample chambers.The method may further comprise providing a plurality of second samplechambers with a second biological material, and transporting the secondbiological material from the second sample chambers into the firstsample chambers by rotating the sample disk about the rotational axis sothat the second biological material passes from the second samplechambers through a plurality of radial fill conduits into the firstsample chambers.

[0007] Various aspects comprise an apparatus for centrifugally loadingand thermally cycling a sample disk. The apparatus can comprise a sampledisk having a plurality of first sample chambers, a plurality of secondsample chambers, and a reservoir for storing a volume of liquid sample.The apparatus may further include means for centrifugally loading theplurality of first sample chambers with liquid sample upon rotation ofthe sample disk about a rotational axis of the sample disk. Theapparatus may further includes means for centrifugally loading theplurality of first sample chambers with a biological material from theplurality of second sample chamber. The apparatus may further include ameans for thermally cycling the plurality of first sample chambers ofthe sample disk.

[0008] Various other aspects comprise an apparatus configured forcontaining samples of biological material during a thermal cyclingoperation. The apparatus may include a microcard configured for rotationabout a rotational axis, a plurality of first sample chambers positionedon the microcard around the rotational axis, and a plurality of secondsample chambers positioned in the microcard around the rotational axis.The second sample chambers may be positioned closer to the rotationalaxis than the first sample chambers. The apparatus may further comprisea plurality of channels formed in the microcard. The plurality ofchannels may comprise a plurality of circumferential channels and aplurality of radial channels. The circumferential channels may bepositioned between adjacent first sample chambers to transport a firstbiological material from a reservoir into the plurality of first samplechambers upon rotation of the microcard about the rotational axis. Theplurality of radial channels may be positioned between correspondingfirst and second sample chambers to transport a second biologicalmaterial from the second sample chambers to the first sample chambersupon a further rotation of the microcard about the rotational axis. Theplurality of first sample chambers may be configured to permit opticaldetection of the biological materials in the first sample chambers.

[0009] It is to be understood that both the foregoing generaldescription and the following description of various embodiments areexemplary and explanatory only and are not restrictive.

BRIEF DESCRIPTION OF THE DRAWINGS

[0010] The accompanying drawings, which are incorporated in andconstitute a part of this specification, illustrate several exemplaryembodiments. In the drawings,

[0011]FIG. 1A is a plan view of an exemplary embodiment of a sample diskaccording to the present teachings, prior to spinning the disk, with afirst biological material in a fill chamber;

[0012]FIG. 1B is a plan view of the sample disk of FIG. 1A aftercentrifugal loading of the first biological material into outer samplechambers of the disk;

[0013]FIG. 1C is a plan view of the sample disk of FIG. 1B, with asecond biological material such as a test sample in inner samplechambers of the disk and with circumferential fill conduits being in ablocked state;

[0014]FIG. 1D is a plan view of the sample disk of FIG. 1C after asecond centrifugal loading operation, and with the circumferential fillconduits and radial fill conduits in a blocked state;

[0015]FIG. 2A is a cross-sectional view along line 2A-2A of FIG. 1A;

[0016]FIG. 2B is a cross-sectional view along line 2B-2B of FIG. 1B;

[0017]FIG. 2C is a cross-sectional view along line 2C-2C of FIG. 1C;

[0018]FIG. 2D is a cross-sectional view along line 2D-2D of FIG. 1D;

[0019]FIG. 3A is a plan view of a sample disk according to anotherembodiment of the present teachings, prior to filling a fill chamberwith a test sample material;

[0020]FIG. 3B is a plan view of the sample disk of FIG. 3A after fillingthe fill chamber with a test sample material; and

[0021]FIG. 3C is a plan view of the sample disk of FIG. 3B, aftercentrifugal loading of the test sample material into the samplechambers, and radial fill conduits in a blocked state.

DESCRIPTION OF VARIOUS EMBODIMENTS

[0022] Reference will now be made to various exemplary embodiments,examples of which are illustrated in the accompanying drawings. Whereverpossible, the same reference numbers are used in the drawings and thedescription to refer to the same or like parts.

[0023] In accordance with various embodiments, a rotatable sample diskconfigured for samples of biological material is provided. In oneaspect, the sample disk includes a fill chamber for storing a firstbiological material, a plurality of first sample chambers positioned inthe sample disk, a plurality of second sample disks, and a plurality ofconduits configured to permit transfer of the first biological materialfrom the fill chamber to the plurality of first sample chambers upon afirst rotation of the sample disk about a rotational axis of the sampledisk.

[0024] Although terms like “horizontal,” “vertical,” “upward,”“downward,” “radial,” and “axial” are used in describing various aspectsof the present teachings, it should be understood that such terms arefor purposes of more easily describing the teachings, and do not limitthe scope of the teachings.

[0025] In various embodiments, such as illustrated in FIGS. 1-2, asample disk 10 is provided. The sample disk 10 may be configured forthermally cycling samples of biological material in a thermal cyclingdevice. The thermal cycling device may be configured to perform nucleicacid amplification on samples of biological material. One common methodof performing nucleic acid amplification of biological samples ispolymerase chain reaction (PCR). Various PCR methods are known in theart, as described in, for example, U.S. Pat. Nos. 5,928,907 and6,015,674 to Woudenberg et al., the complete disclosures of which arehereby incorporated by reference for any purpose. Other methods ofnucleic acid amplification include, for example, ligase chain reaction,oligonucleotide ligations assay, and hybridization assay. These andother methods are described in greater detail in U.S. Pat. Nos.5,928,907 and 6,015,674.

[0026] In various embodiments, the sample disk may be used in a thermalcycling device that performs real-time detection of the nucleic acidamplification of the samples in the sample disk during thermal cycling.Real-time detection systems are known in the art, as also described ingreater detail in, for example, U.S. Pat. Nos. 5,928,907 and 6,015,674to Woudenberg et al., incorporated herein above. During real-timedetection, various characteristics of the samples are detected duringthe thermal cycling in a manner known in the art. Real-time detectionpermits more accurate and efficient detection and monitoring of thesamples during the nucleic acid amplification process. Alternatively,the sample disk may be used in a thermal cycling device that performsendpoint detection of the nucleic acid amplification of the samples. Onetype of detection apparatus that may be used with the present teachingsfor either real-time or endpoint detection is the LightCycler™Instrument manufactured by Roche Molecular Biochemicals. Another type ofdetection apparatus includes a single LED sensor for detecting thecharacteristics of the samples as the sample disk rotates about arotational axis. Several other types of detection apparatus are shown inWO 02/00347A2 to Bedingham et al., the complete disclosure of which ishereby incorporated by reference for any purpose.

[0027] The sample disk may be configured to contact a sample block forthermally cycling the biological materials in the sample chambers of thesample disk. The sample block may be operatively connected to atemperature control unit programmed to raise and lower the temperatureof the sample block according to a user-defined profile. For example, invarious embodiments, a user may supply data defining time andtemperature parameters of the desired PCR protocol to a control computerthat causes a central processing unit (CPU) of the temperature controlunit to control thermal cycling of the sample block. Severalnon-limiting examples of suitable temperature control units for raisingand lowering the temperature of a sample block for a microcard or othersample-holding member are described in U.S. Pat. No. 5,656,493 to Mulliset al. and U.S. Pat. No. 5,475,610 to Atwood et al., the disclosures ofwhich are both hereby incorporated by reference for any purpose.

[0028] In one embodiment, the rotatable sample disk comprises at leastone fill chamber on the rotatable sample disk, a plurality of firstsample chambers, a plurality of second sample chambers, and a pluralityof fill conduits. One embodiment of a sample disk of the presentteachings is shown in FIGS. 1-2. As embodied herein and shown in FIGS.1-2, the rotatable sample disk is a microcard or sample tray generallydesignated by reference number 10. The sample disk is generallyrotatable about a rotational axis 12. The rotatable sample disk 10 isshown as being a circular plate, however, it is understood that thesample disk may be any other suitable shape such as rectangular orsquare. A circular shape is shown merely because a circular shape willtypically minimize the amount of space taken up by the sample disk as itrotates about rotational axis 12.

[0029] As shown in FIGS. 1-2, particularly FIG. 2A, the rotatable sampledisk may include a first layer 14 and second layer 16. For purposes ofconvenience, the first layer may be referred to as the “top layer” andthe second layer may be referred to as the “bottom layer.” As shown forexample in FIG. 2A, the first layer 14 includes a top surface 18 andbottom surface 20. The second layer 16 generally includes a top surface22 and a bottom surface 24. The first and second layers may be made outof any suitable material or materials. In a typical embodiment, thefirst layer 14 is made of a polymeric material such as polypropylene andthe second layer 16 is made out of a metal such as metal foil.Alternatively, both the first layer 14 and the second layer 16 may bemade out of a polymeric material. In another embodiment, the first layeris made out of polypropylene and the second layer is made out of lexan.Other suitable polymers include polyester, polycarbonate, andpolyethylene.

[0030] In the embodiment shown, the first layer 14 includes all of thefeatures of the sample chambers, fill conduits, and fill chambers in apolymeric sheet that has been molded, vacuum formed, pressure formed,compression molded, or otherwise processed. The second layer 16 isprovided as a substantially flat plate that is attached to the firstlayer 14 to complete formation of the features of the sample chambers,fill conduits, and fill conduits. It should be understood that thefeatures may be provided in both layers of the sample disk. It may bedesired that the first and second layer are made out of PCR-compatiblematerials. It may also be desirable that the materials selected for thefirst and second layer exhibit good water barrier properties.

[0031] A variety of methods of forming the layers and methods ofadhering the two layers together are described in, for example, WO02/01180A2 to Bedingham et al., the complete disclosure of which ishereby incorporated by reference for any purpose, and WO 02/00347A2 toBedingham et al., incorporated herein above. The structure of the firstand second layers will be described in greater detail below, as thestructure of the first and second layers define the sample chambers,fill chambers, and fill conduits that comprise sample disk 10.

[0032] In various embodiments, the sample disk includes at least onefill chamber for storing a first biological material, and a plurality offirst sample chambers. As embodied herein and shown in FIGS. 1A-1D, thesample disk includes a fill chamber 28 positioned on the upper layer 14of the sample disk, and a plurality of first sample chambers 40 (alsoreferred to as “outer sample chambers”). The fill chamber of oneembodiment of the present teachings serves as a reservoir for storingthe first biological material prior to the sample disk being rotated tocentrifugally load the first biological material into the outer samplechambers.

[0033] In the embodiment shown in FIGS. 1-2, the first biologicalmaterial would typically be a reagent, particularly an analyte-specificreagent. Analyte-specific reagents are well-known in the art. It shouldbe understood that the first biological material may be any other typeof suitable biological material, such as a test sample material, insteadof a reagent. For purposes of conveniently describing the embodiment ofFIGS. 1-2, the first biological material will be described as a reagent.In the embodiment shown in FIGS. 1-2, the user can select an appropriatereagent or other biological material, thereby providing more flexibilitycompared to testing devices in which the reagents are pre-programmedinto the testing device. If the sample disk provides for a single“primary” fill chamber 28, one reagent may be used in a single sampledisk.

[0034] The fill chamber may have any type of shape suitable for storinga liquid. In the example shown in FIGS. 1A-1D, the fill chamber 28 isshown as being generally oval, however any other suitable shape isacceptable. The volume of the fill chamber can range from quite large tovery small, depending on the desired amount of reagent (or other firstbiological material) for each of the outer sample chambers 40 into whichit will be centrifugally loaded in a manner described below. Typically,the total amount of reagent placed in the fill chamber will bepredetermined prior to entry of the reagent into the fill chamber 28.The amount of volume may be calculated based on the amount of reagentdesired in each sample chamber, multiplied by the total number of samplechambers on the sample disk. By way of example only, in an embodiment inwhich there are seventy-two sample chambers 28, the predetermined amountof reagent to be inserted into the fill chamber 28 may be seventy-twotimes the amount of reagent desired in each outer sample chamber. Forexample, in a scenario in which it is desired that each of theseventy-two outer sample chambers eventually contain approximately 5 μlof reagent, then the approximate total volume of the fill chamber wouldbe approximately 360 μl. The desired amount of reagent can greatly varyhowever, depending on a large number of factors.

[0035] In various embodiments, the fill chamber may include an orificefor permitting loading of the first biological material into the fillchamber. As shown in FIGS. 1A-1D, orifice 30 may be provided on theoutside of the fill chamber 28. The orifice 30 is typically sized inorder to permit pipetting of the first biological material, such as areagent, into the fill chamber. Alternatively, the fill chamber may befilled by any other acceptable method for inserting a first biologicalmaterial such as a reagent into a reservoir. It should be understoodthat although the drawings only illustrate a single fill chamber 28, itis easily understood that the sample disk could have any number of“primary” fill chambers. The fill chambers could be positioned aroundthe rotational axis, typically in a concentric and evenly spaced mannerin order to promote a uniform distribution of the first biologicalmaterial into the sample chambers.

[0036] In various embodiments, the sample disk includes a plurality offirst sample chambers, a plurality of second sample chambers, and aplurality of fill conduits. In the embodiment shown in FIGS. 1-2, thesample disk includes a plurality of first sample chambers (or “outersample chambers”) 40 and a plurality of second sample chambers (or“inner sample chambers”) 48. As shown for example in FIG. 1A, theplurality of outer sample chambers 40 are positioned concentricallyabout the rotational axis of the 12. It is also contemplated that theouter sample chambers may be positioned non-concentrically, however itis typically desired to have the outer sample chambers positionedconcentrically to enhance uniform volumes of biological material in eachof the outer sample chambers. The outer sample chambers 40 may beequally spaced from one another as shown in FIGS. 1A and 1B, or thespacing may be varied.

[0037] The sample chambers may have any shape suitable for thermalcycling. In the embodiment shown in FIGS. 1-2, the outer sample chambers40 are cylindrical with flat top surface 44, however, any other knownshape is also suitable. In a typical system, light may be transmittedthrough the top surface of the outer sample chambers during detection ofthe characteristics of the biological material in the sample chamber. Asbest seen in FIG. 2A, outer sample chamber 40 is formed by a raised flattop surface 44 of first layer 14 that creates a space between the firstlayer and the top surface 22 of second layer 16. The raised flat topsurface 44 may be formed by any known method. The outer sample chamber40 defines a volume for storing biological materials.

[0038] In the embodiment shown in FIG. 1A, a total of seventy-two (72)outer sample chambers are included on the sample disk, however it ispossible to use anywhere from one to at least several thousand outersample chambers. The outer sample chambers are preferably configured tobe PCR-compatible, and typically have a surface such as top surface 44through which an optical detection system (not shown) can detect thecharacteristics of sample materials stored in the sample chambers. Theconcept of sample chambers is known in the art. In a more typicalembodiment, the size of the sample chambers may vary from 0.1 μl toseveral thousand μl. In a more typical embodiment such as shown in FIG.1, the outer sample chambers 40 are configured to have a volume ofapproximately 10 μl. It should be understood that this volume is forpurposes of example only. In some instances, it may be desirable to havesmaller volumes in order to reduce the amount of reagent and samplematerial required to load the sample disk. In other instances, it may bedesirable to have a greater volume. In various embodiments, the chambersare configured to hold no greater than 1,000 μl. In other embodiments,the chambers are configured to hold no more than 200 μl, no more than100 μl, no more than 50 μl, or no more than 0.5 μl.

[0039] In accordance with various embodiments, the sample disk includesa plurality of fill conduits configured to permit transfer of a firstbiological material from the fill chamber to the plurality of firstsample chambers upon rotation of the sample disk about the rotationalaxis. As embodied herein and shown in FIGS. 1A and 1B, the plurality offill conduits includes a plurality of circumferential fill conduits 42positioned between adjacent outer sample chambers 40. In the embodimentshown in FIG. 1A, the circumferential fill conduits 42 are positionedconcentrically about the rotational axis 12 at a fixed diameter. Itshould be understood that the circumferential fill conduits do notnecessarily need to be concentrically spaced from the rotational axis12. In FIGS. 1A and 1B, the circumferential fill conduits 42 are shownbisecting the center of the sides of each of the outer sample chambers40. The circumferential fill conduits are designed to permit fluidcommunication between adjacent outer sample chambers. In the embodimentshown, the circumferential fill conduits 42 are defined by featuresformed in the first layer 14 that create a space with the top surface 22of the second layer 16. The features in the first layer may be formed byany known processing method such as, but not limited to, molding, vacuumforming, pressure forming, and compression molding. In variousembodiments, the fill conduits (or channels) described herein may have arange of sizes. In various embodiments, such conduits have at least onecross-sectional dimension, e.g., width, depth, or diameter, of between 1to 750 micrometers. In various other embodiments, such conduits have atleast one cross-sectional dimension of from between 10 to 500micrometers, or from between 50 to 250 micrometers.

[0040] In various embodiments, the sample disk comprises a primary fillconduit extending from the fill chamber to the circumferential fillconduits and/or outer sample chambers. As shown in FIGS. 1A-1D, aprimary fill conduit 46 may extend between the fill chamber 28 to one ofthe circumferential fill conduits 42. In one embodiment, the primaryfill conduit 46 extends radially in order to transport the reagent inthe fill chamber to the circumferential fill conduit 42 upon rotation ofthe sample disk about rotational axis 12. The primary fill conduit maybe sized to prevent the first biological material, typically reagent R,from passing through it while the sample disk is stationary. If theprimary fill conduits are an appropriate size and shape, the surfacetension of the interior surface of the conduit will prevent the flow ofthe reagent through the primary fill conduit when the sample disk is ata resting position (or rotating at a speed below a predetermined speedat which the reagent will begin to flow due to centrifugal force).

[0041] It should be understood that the primary fill conduit need not becompletely radial in order to transport the reagent. Likewise, it shouldalso be understood that, in some embodiments, the primary fill conduitcould be eliminated by moving the fill chamber closer to or adjacent thecircumferential fill conduit. Although only one primary fill conduit 46is shown in FIGS. 1A-1D, it is contemplated that several fill conduitscould be used, particularly in embodiments having a plurality of fillchambers. For example, it is conceivable to have the same number of fillchambers as outer sample chambers. In such an embodiment, each fillchamber may have an individual primary fill conduit. With an embodimenthaving seventy-two outer sample chambers, a total of seventy-two fillchambers and seventy-two primary fill conduits might be provided. It isof course contemplated that a smaller number of fill chambers andprimary fill conduits could also be used.

[0042] It should also be understood that several sets of fill chambersand outer sample chambers may be provided. In one embodiment, each setof fill chambers and one or more sample chambers could use separatesamples to be tested. This would allow for a large amount of samples tobe tested on a single sample disk. In another embodiment, each fillchamber could be ganged with one or more outer sample chambers. Itshould also be understood that a plurality of disks could be stackedtogether.

[0043] The sample disk may further include a plurality of inner samplechambers positioned radially inside of the outer sample chambers. In theembodiment shown for example in FIGS. 1A and 2A, the inner samplechambers 48 are similar in shape to the outer sample chambers 40. Theinner sample chambers 48 however may have any shape suitable for storinga second biological material. Inner sample chambers 48 may be positionedcloser to the rotational axis 12 than the outer sample chambers 40,i.e., radially inside of the outer sample chambers. In the embodimentshown in FIGS. 1-2, the inner sample chambers 48 may be used to store asecond biological material, typically the sample material to be tested,after a first biological material such as a reagent has beencentrifugally loaded from the fill chamber to the outer sample chambers40. After the reagent has been centrifugally loaded into the outersample chambers as shown in FIGS. 1B and 2B, the inner sample chambers48 may be filled with a sample to be tested.

[0044] As shown in FIG. 1A, in a typical arrangement, an equal number ofinner sample chambers and outer sample chambers are provided. In theexample shown in FIG. 1A, the sample disk includes seventy-two innersample chambers 48 and seventy-two outer sample chambers 40. It isunderstood that any other number of inner sample chambers may beprovided. In the embodiment shown, the inner sample chambers 48 areapproximately the same size as the outer sample chambers 40, howeverthis can be varied depending on the specific application. For example,in a sample disk configuration in which it is desired to use a largeamounts of reagent and a small amount of test sample material, it may bedesired to have inner sample chambers that are smaller than the outersample chambers. Likewise, in the situation where it is not desired tocompletely fill the outer sample chambers, it may also be desired tohave inner sample chambers that are smaller than the outer samplechambers.

[0045] In accordance with various embodiments, radial fill conduits maybe provided between the second sample chambers and the first samplechambers. In the embodiment shown in FIG. 1, the second or inner samplechambers 48 and first or outer sample chambers 40 are connected byradial fill conduits 50. As shown in FIG. 1A, radial fill conduit 50extends in a radial direction with respect to rotational axis 12. FIG.2A illustrates a cross-section along line 2A-2A of FIG. 1A which passesthrough radial fill conduit 50. As can be seen in FIG. 2A, radial fillconduit is configured to permit fluid communication between the innersample chamber 48 and outer sample chamber 40. The radial fill conduitis defined by a raised portion 52 in the first layer 14 of the sampledisk, and a top surface 22 of the second layer 16 of the sample disk.The raised portion is formed in the first layer 14 by any known method.The radial fill conduits are typically sized in depth so that fluid onlypasses through the conduit upon a centrifugual force being imparted onthe sample disk.

[0046] The radial fill conduits 50 are shown having approximately thesame size as the circumferential fill conduits 42, however the size andshape of the radial fill conduits may be varied. As will be describedlater, the radial fill conduits permit transfer of a second biologicalmaterial from the inner sample chambers 48 to the outer sample chambers40 upon rotation of the sample disk. In the embodiment described herein,the loading of the second biological material, e.g., sample to betested, into the outer sample chambers typically occurs after the firstbiological material, e.g., reagent, has already been pre-loaded into theouter sample chambers. This will be discussed in greater detail in thedescription of the operation of the sample disk.

[0047] In various embodiments, the inner sample chambers 48 may includean orifice for permitting loading of the second biological material intothe inner sample chamber. As shown in FIGS. 1A and 2A, for example, anorifice 54 may be formed in the top surface 56 of inner sample chamber48. The orifice allows for manual or automatic loading of the secondbiological material, typically the sample test material, into the innersample chamber 48. A typical method of loading the sample test materialis pipetting. Other methods may also be utilized however. Instead of anorifice, any other known type of structure for permitting entry of aliquid into a reservoir may also be used.

[0048] In various embodiments, a means for selectively blocking thepassage of the first biological material through the circumferentialfill conduits is provided. The blocking of the passage of the firstbiological material, typically a reagent, through the circumferentialfill conduits is particularly useful after the reagent has already beencentrifugally loaded from the primary fill conduit into the outer samplechambers. The blocking assists in preventing cross-contamination betweenadjacent outer sample chambers. In the embodiment shown in FIG. 1C, thefirst layer 14 of the sample disk may be physically deformed at position60 by a staking device in order to block or occlude the circumferentialfill conduits 42. The staking device may be any device that isconfigured for physically deforming the circumferential fill conduit,such as a knife edge. Alternatively, the second layer 16 may bephysically deformed in order to block or occlude the circumferentialfill conduits. The materials of the sample disk are typically selectedso that staking may effectively occur.

[0049] It should be understood, however, that the complete sealing oroccluding of the circumferential fill conduits may not be required. Forexample, it may only be required that the deformation restrict flow,migration or diffusion through a conduit or fluid passageway sufficientto provide the desired isolation of adjacent outer sample chambers. Asused in connection with the present teachings, “blocking” or “occlusion”or “closing” will include both partial blocking and complete blocking.

[0050] In order to promote more effective blocking, it may be desired touse any other known means for blocking a conduit. For example, it may beuseful to use adhesives on either or both of the first and second layerin order to promote sealing of the conduit after the layers aredeformed. Instead of physical deformation, the means for blocking mayalso comprise any other type of melting, bonding, and welding in orderto block off the circumferential fill conduit. A number of suitablemethods of blocking or occluding a conduit of a microcard are describedin WO 02/01180A2 to Bedingham et al., incorporated by reference above.

[0051] In various embodiments, a means for selectively blocking thepassage of the first and second biological material from the outersample chambers to the inner sample chambers is provided. The means forblocking is utilized after the biological sample material to be tested,shown as S in FIG. 1C, has been centrifugally loaded from the innersample chambers 48 into the outer sample chambers 40. When the sample tobe tested is centrifugally loaded into the outer sample chamber 40 itmixes with the reagent, shown as R, which was previously loaded into theouter sample chambers 40 as shown in FIGS. 1B and 2B. After the sampleto be tested is loaded into the outer sample chambers, it is desiredmaintain the sample and reagent in the outer sample chambers. A numberof different methods and structures may be used to block the radial fillconduits in order to maintain the sample and reagent in the outer samplechambers.

[0052] For example, FIGS. 1D and 2D show an embodiment in which thefirst layer 14 of the sample disk is physically deformed at position 70by a staking device, such as a knife edge, in order to block passage ofthe liquids out of the outer sample chambers. FIG. 2D shows thephysically deformed portion 70 of the first layer 14 that has beenpressed downward by a staking apparatus in order to block or occlude thepassage of liquid through radial fill conduit 50. This blocking isuseful so that a constant volume of material is maintained in each ofthe outer sample chambers 40 during thermal cycling and detection of thesample chambers. This will be discussed in greater detail in thedescription of the operation of the sample disk.

[0053] An example of the operation of the sample disk for the embodimentof FIGS. 1-2 is described below. In the first step of the operation, asample disk is provided. As shown in FIG. 1A, the sample disk 10 of oneembodiment includes, among other things, a fill chamber 28, a primaryfill conduit 46, a plurality of outer sample chambers 40, a plurality ofinner sample chambers 48, a plurality of circumferential fill conduits42, and a plurality of radial fill conduits 50. The sample disk 10 istypically placed on the drive shaft of a centrifuge in a thermal cyclingdevice (not shown) so that the sample disk may rotate about rotationalaxis 12.

[0054] Next, a first biological material may be loaded into the fillchamber 28 positioned on the sample disk. As previously discussed, thefirst biological material in the embodiment of FIGS. 1-2 is typically areagent. For the sake of ease of discussion, the first biologicalmaterial will be referred to as a reagent. It should be understoodhowever, that the first biological material may instead be the sample tobe tested. The reagent is labeled R in FIG. 1A. The reagent may beloaded into the fill chamber 30 by any suitable method. One method is topipette the reagent through orifice 30 positioned on top of the fillchamber 28. In the position shown in FIG. 1A, the reagent fills the fillchamber 28 but does not pass along the primary fill conduit 46 due tothe surface tension of the reagent on the primary fill conduit. As shownin FIG. 2A, the outside sample chamber 40 and inner sample chamber 48are initially empty.

[0055] After a predetermined amount of reagent has been loaded into thefill chamber 28, the sample disk 10 is rotated about the rotational axisby the centrifuge (not shown). Upon reaching a certain rotational speed,the centrifugal force will cause the reagent R to flow through theprimary fill conduit 46 and through the circumferential fill conduits 42into the outer sample chambers 40 until the reagent is evenlydistributed throughout the outer sample chambers as shown in FIGS. 1Band 2B. In one exemplary embodiment, the reagent fills approximatelyhalf of each outer sample chamber, as shown in FIGS. 1B and 2B. Becauseof the centrifugal force, the reagent will fill the half of the outersample chamber farthest from rotational axis 12. It should be understoodthat a greater amount of reagent may be used. A smaller amount ofreagent may also be used. If a smaller amount of reagent is used, thesample disk may be modified so that the circumferential fill conduits 42are positioned farther from the rotational axis 12 than shown in FIGS.1-2. The circumferential fill channels would be moved closer to theouter edge of the outer sample chambers 40.

[0056] It should of course be understood that the centrifugal force tocause loading of the outer sample chambers may be created by any numberof rotations, including less than a full rotation.

[0057] The circumferential fill conduits 42 shown in FIGS. 1-2 may nowbe staked by any known method such as physical deformation of the firstlayer 14 of the sample block to occlude the passage of fluid through thecircumferential fill conduits 42. FIGS. 1C and 2C show thecircumferential fill conduits 42 being staked at position 60 to preventfluid communication between adjacent outer sample chambers 40.

[0058] After the circumferential fill conduits 42 have been staked, asecond biological material, typically a sample S to be tested, may beloaded into the inner sample chambers 48. It should be understood thatthe second biological material may instead be a reagent or other type ofbiological material. In one embodiment, the test sample S is insertedinto some, if not all, of inner sample chambers 48 shown in FIGS. 1C and2C through orifice 54 on the top surface 56 of the inner sample chamber48. The surface tension of the sample prevents the sample from initiallyflowing through the radial fill conduit 50 into the outer sample chamber40.

[0059] After the sample S has been loaded into the inner sample chambers48, and the circumferential fill conduits 42 have been staked, both asshown in FIGS. 1C and 2C, the sample disk is rotated again about therotational axis 12. Upon rotation of the sample disk at a certain speed,the sample S to be tested will be urged by centrifugal force to flowfrom the inner sample chambers 48 through the radial fill conduits 50into the outer sample chambers 40. The sample to be tested flows intothe outer sample chambers 40 and mixes with the reagent R to form afinal test material labeled F in FIGS. 1D and 2D. After the sample to betested has been loaded into the outer sample chambers, the radial fillconduits 50 are staked to maintain the final test material F (comprisingthe reagent and the sample to be tested) in the outer sample chambers40. As shown in FIG. 2D, the radial fill conduit may be staked, in oneembodiment, by physically deforming a portion of the top surface of thefirst layer 14 in a region intersecting the radial fill conduit. Theportion of the top surface that is deformed is labeled as referencenumber 70 in FIG. 2D. After the radial fill conduit is sufficientlyblocked, the sample disk may be further rotated and thermally cycled.

[0060] During rotation and thermal cycling of the sample block, in oneembodiment, the optical characteristics of the final sample F can bedetected by an optical detection system. In one embodiment, the opticaldetection system is similar to the LightCycler™ system of Roche.

[0061] In this embodiment, the sample disk may be maintained about asingle rotational axis for the processes of filling the sample chambers,thermally cycling the samples, and optically detecting the samples. Bysuch an operation, the cost and time spent loading the sample chambersand thermally cycling the sample chambers may be minimized. Moreover, byproviding a sample disk that can be loaded by rotating about a singlerotational axis, it is possible to provide an integrated centrifuge andthermal cycling device. Such an integrated centrifuge and thermalcycling device could conceivably be a portable apparatus that can beuseful for point of service analysis.

[0062] As is clear from the above description, the present teachingsinclude methods of centrifugally loading a plurality of sample chambersin a sample disk. The method may comprise providing a sample disk with afill chamber, a plurality of first sample chambers, and a plurality ofsecond sample chambers. The method may comprise inserting a firstbiological material into the fill chamber. The method may furthercomprise loading the plurality of first sample chambers with a firstbiological material by rotating the sample disk about a rotational axisso that the first biological material passes from the fill chambertravels through a plurality of circumferential fill conduits connectingthe fill chamber with the first sample chambers. The plurality ofcircumferential fill conduits may be positioned between adjacent firstsample chambers. The method may further comprise providing a pluralityof second sample chambers with a second biological material, andtransporting the second biological material from the second samplechambers into the first sample chambers by rotating the sample diskabout the rotational axis so that the second biological material passesfrom the second sample chambers through a plurality of radial fillconduits into the first sample chambers.

[0063] In accordance with further various embodiments, the rotatablesample disk configured for thermally cycling samples of biologicalmaterial comprises a sample disk with a fill chamber, a primary fillconduit, a circumferential fill conduit, a plurality of radial fillconduits, and a plurality of sample chambers. In these variousembodiments, a biological material such as a reagent may be insertedinto the sample chambers prior to a sample to be tested beingtransferred from the fill chamber into the sample chambers bycentrifugal force. In various embodiments, the sample disk may be filledwith biological material, thermally cycled, and optically detected,without removing the sample disk from a rotational axis.

[0064] Further various embodiments of the sample disk contemplatestructure such as shown in FIGS. 3A-3C. The sample disk is generallydesignated by the reference number 100 in FIGS. 3A-3C. To the extentthat any of the following structure is similar to the structuredescribed above for the embodiments shown in FIGS. 1-2, a detaileddescription will not be repeated. In the embodiment shown in FIGS.3A-3C, the sample disk is rotatable about a rotational axis. The sampledisk may be formed in a similar manner as discussed for the embodimentsshown in FIGS. 1-2.

[0065] As embodied herein and shown in FIGS. 3A-3C, sample disk 100 mayinclude a fill chamber 110 for storing a biological material and anorifice 112 for permitting loading of the biological material into thefill chamber. The fill chamber 110 may be similar to the one describedfor the FIGS. 1-2 embodiments, or it may be completely different. Thefill chamber serves as a reservoir for storing the biological materialprior to centrifugal loading to transfer the biological material intosample chambers.

[0066] In various embodiments, the sample disk includes a plurality ofsample chambers 120 in fluid communication with the fill chamber 110through a primary fill conduit 122, a circumferential fill conduit 124,and a plurality of radial fill conduits 130. The sample chambers 120 maybe any type of sample chamber suitable for thermal cycling and opticaldetection of the characteristics of the sample to be contained therein.Suitable sample chambers are described in relation to the embodiment ofFIGS. 1-2. Although the figure show seventy-two sample chambers, anynumber of sample chambers from one to several thousand may be used withthe embodiment of FIGS. 3A-3C.

[0067] The sample disk 100 of FIGS. 3A-3C will typically, but notalways, have a reagent pre-inserted into each sample chamber 120. Thereagent is indicated by reference letter R in FIGS. 3A and 3B. This isreferred to as a pre-programmed sample disk. Alternatively, the sampledisk may have a sample to be tested pre-inserted into each samplechamber instead of a reagent. In one typical embodiment, all of thesample chambers include at least one reagent. In other variations, onlysome of the sample chambers contain a reagent. In still othervariations, none of the sample chambers contain a reagent. In a typicalapplication, the reagent is dried into the sample chambers prior to anytest sample material being allowed to enter the sample chambers. Methodsof inserting and fixing the reagents to the inside of the samplechambers are known in the art, and will not be described here. As statedabove, the reagent is optional.

[0068] The sample disk may further include a primary fill conduit 122extending radially from the primary fill conduit. The primary fillconduit 122 may be essentially identical to any of those described abovefor the embodiments of FIGS. 1-2. As discussed above for FIGS. 1-2, aprimary fill conduit may not be necessary in some variations.

[0069] In accordance with various embodiments, the primary fill conduitand/or fill chamber may be in fluid communication with a circumferentialfill conduit. As shown in FIGS. 3A-3C, the sample disk includes acircumferential fill conduit 124. The circumferential fill conduit shownin FIGS. 3A-3C is a continuous conduit with a constant diameter from therotational axis 102. It should be contemplated that the sample disk maycontain several such fill conduits. Moreover, the circumferential fillconduit need not be concentric or continuous. In the embodiment shown inFIGS. 3A-3C, the circumferential fill conduit is positioned radiallyinside of the sample chambers 120.

[0070] In accordance with various embodiments, the circumferential fillconduit may be in fluid communication with a plurality of radial fillconduits extending radially from the circumferential fill conduit. Asshown for example in FIGS. 3A-3C, the sample disk includes a pluralityof radial fill conduits 126 extending radially from the circumferentialfill conduit 124 into the sample chambers 120. The radial fill conduits126 branch out from the circumferential fill conduit in a radialdirection relative to the rotational axis. In a typical embodiment, thesample disk has an equal number of radial fill conduits and samplechambers. It should be understood that the features of the sample disksuch as the radial fill conduits, circumferential fill conduit, andprimary fill conduit may be formed by any of the methods discussed forthe same in the embodiment of FIGS. 1-2.

[0071] In various embodiments, such as shown in FIGS. 3A-3C, a means forselectively blocking the flow of the first and second biologicalmaterials from the sample chambers to the circumferential fill conduitis provided. As shown in FIG. 3C, each radial fill conduit 126 may bestaked to prevent the flow of the final sample to be tested, referred toas F, from the sample chambers into the circumferential fill conduit124. The position along which the staking may be performed is generallyindicated by reference number 130. The staking can be done by any of themethods discussed above for the embodiment shown in FIGS. 1-2.

[0072] The sample disk may also contain an area 130 where informationabout the sample disk or its contents may be stored. Such informationmay be in the form of a bar code, as shown in FIGS. 3A-3D, writteninformation, or any other form suitable for displaying characteristicsof the sample disk or the samples contained therein.

[0073] An example of the operation of the sample disk for the embodimentof FIGS. 3A-3C is described below. To the extent that the followingoperation is similar to the operation described above for the embodimentshown in FIGS. 1-2, a detailed description of the operation will not berepeated. As shown in FIG. 3A, the sample disk 100 of one embodimentincludes, among other things, a fill chamber 110, a primary fill conduit122, a plurality of sample chambers 120, a circumferential fill conduit124, and a plurality of radial fill conduits 126. The sample disk may beused with a thermal cycling device identical to that described for theFIG. 1-2 embodiment above. The sample disk shown in FIG. 3A typicallyincludes reagents in some, if not all, of the chambers. These reagentswould typically be inserted and dried into the sample chambers prior tothe user inserting the sample disk into the thermal cycling device. Thefill chamber 110 is empty in FIG. 3A.

[0074] Next, as shown in FIG. 3B, a sample S to be tested may be loadedinto the fill chamber 110 of the sample disk 100. In a typically method,the sample to be tested is pipetted through orifice 112 into the fillchamber 110. The amount of sample to be loaded into the fill chamber maybe predetermined by calculating the total amount of amount of sample tobe tested that is desired in each of the sample chambers 120 and thenmultiplying by the number of sample chambers. As previously discussedfor FIGS. 1-2, the liquid in the fill chamber will typically remainwithin the fill chamber due to surface tension of the liquid, until thesample disk is rotated to a sufficient speed.

[0075] Next, the sample disk may be rotated by the centrifuge so thatthe sample S to be tested is urged by centrifugal force to flow from thefill chamber 110 through the primary fill conduit 122 andcircumferential fill conduit 124 into each of the radial fill conduits126. The sample then travels into the sample chambers 120. In oneembodiment, the sample is evenly distributed among the sample chambersso that each sample chamber has an essentially identical amount ofsample liquid. The final material to be tested (comprising a mixture ofthe reagent R and the sample S) is indicated by reference letter F inFIG. 3C.

[0076] After the sample S to be tested has been loaded into the samplechambers to join the already inserted reagents R to form final testmaterial F, the sample disk may be staked. As shown in FIG. 3C, theradial fill conduits 126 may be staked, in one embodiment, by physicallydeforming portion 130 of the top layer of the sample disk in a mannersimilar to that performed on radial fill conduits 50 in the FIG. 1-2embodiments. After the radial fill conduit is sufficiently blocked, thesample disk may be further rotated and thermally cycled. During or afterthermal cycling, the optical characteristics of the final sample F maybe detected by an optical detections system. In one embodiment, theoptical detections sytem is similar to the LightCycler™ system of RocheMolecular Biochemicals.

[0077] As is clear from the above description, the present teachings mayalso include a method of centrifugally loading and thermally cycling aplurality of sample chambers on a sample disk.

[0078] It will be apparent to those skilled in the art that variousmodifications and variations can be made to the structure and methodsdescribed above. Thus, it should be understood that the presentteachings are not limited to the examples discussed in thespecification. Rather, the present teachings are intended to covermodifications and variations.

What is claimed is:
 1. A rotatable sample disk configured for samples ofbiological material, comprising: a fill chamber for storing a firstbiological material, the fill chamber configured for rotation on thesample disk about a rotational axis; a plurality of first samplechambers positioned in the sample disk farther from the rotational axisthan the fill chamber; a plurality of second sample chambers, eachsecond sample chamber configured to permit fluid communication with arespective first sample chamber, the second sample chambers positionedcloser to the rotational axis than the first sample chambers; and aplurality of circumferential fill conduits positioned adjacent theplurality of first sample chambers, the plurality of circumferentialfill conduits configured to permit transfer of the first biologicalmaterial from the fill chamber to the plurality of first sample chambersupon a first rotation of the sample disk about the rotational axis. 2.The rotatable sample disk of claim 1, further comprising a plurality ofradial fill conduits, each radial fill conduit being positioned betweena second sample chamber and a first sample chamber to permit fluidcommunication between the second sample chamber and the first samplechamber.
 3. The rotatable sample disk of claim 2, wherein eachcircumferential fill conduit is positioned between adjacent first samplechambers to permit fluid communication between the adjacent first samplechambers.
 4. The rotatable sample disk of claim 3, the plurality ofcircumferential fill conduits configured so that upon rotation of thesample disk the first biological material contained in the fill chamberis permitted to flow from the fill chamber into the first samplechambers via the circumferential fill conduits.
 5. The rotatable sampledisk of claim 4, further comprising means for selectively blocking thepassage of the first biological material through the circumferentialfill conduits after the first biological material is permitted to flowinto the first sample chambers.
 6. The rotatable sample disk of claim 5,the means for blocking the passage of the first biological materialcomprising an elastically deformable region configured such that uponpressing a staking device against a surface of the sample disk in theelastically deformable region the circumferential fill conduit betweenadjacent first sample chambers is closed off.
 7. The rotatable sampledisk of claim 4, wherein the fill chamber includes an orifice forpermitting entry of the first biological material into the fill chamber.8. The rotatable sample disk of claim 4, wherein the second samplechambers each include an orifice for permitting entry of a secondbiological material into the second sample chambers.
 9. The rotatablesample disk of claim 2, wherein the radial fill conduits are configuredto permit fluid communication of a second biological material from asecond sample chamber to a first sample chamber upon rotation of thesample disk.
 10. The rotatable sample disk of claim 9, furthercomprising means for selectively blocking the passage of the first andsecond biological materials from the first sample chambers to the secondsample chambers.
 11. The rotatable sample disk of claim 10, the meansfor selectively blocking the fluid communication of the first and secondbiological materials from the first sample chambers to the second samplechambers comprising a surface of the sample disk that has beenphysically deformed to close off the radial fill conduits between thefirst sample chambers and the second sample chambers.
 12. The rotatablesample disk of claim 10, the means for selectively blocking the fluidcommunication of the first and second biological materials from thefirst sample chambers to the second sample chambers comprising a one-wayvalve positioned in the radial fill conduits.
 13. The rotatable sampledisk of claim 1, wherein the sample disk comprises a substantially flatcircular plate.
 14. The rotatable sample disk of claim 1, wherein thesample disk is thermally connected to a sample block of a thermalcycling device.
 15. The rotatable sample disk of claim 14, wherein thesample block is operatively connected to a temperature control unit forraising and lowering the temperature of the sample block according to auser-defined profile.
 16. Method of loading a plurality of samplechambers in a sample disk, comprising: providing a sample disk with afill chamber, a plurality of first sample chambers, and a plurality ofsecond sample chambers; loading the plurality of first sample chamberswith a first biological material by rotating the sample disk about arotational axis so that a first biological material in the fill chambertravels through a plurality of circumferential fill conduits connectingthe fill chamber with the first sample chambers, the plurality ofcircumferential fill conduits being positioned between adjacent firstsample chambers; providing a plurality of second sample chambers with asecond biological material; and transporting the second biologicalmaterial from the second sample chambers into the first sample chambersby rotating the sample disk about the rotational axis so that the secondbiological material passes from the second sample chambers through aplurality of radial fill conduits into the first sample chambers. 17.The method of claim 16, wherein the step of providing a plurality ofsecond sample chambers with a second biological material comprisespipetting the second biological material into the second sample chambersthrough orifices in the second sample chambers.
 18. The method of claim16, further comprising, between the step of loading the plurality ofsample chambers with a first biological material and the step ofproviding a plurality of second sample chambers with a second biologicalmaterial, performing the step of isolating adjacent first samplechambers from one another.
 19. The method of claim 18, wherein the stepof isolating adjacent first sample chambers is performed by deformingthe circumferential fill conduits so that liquid cannot pass betweenadjacent first sample chambers.
 20. The method of claim 18, furthercomprising, after the step of transporting the second biologicalmaterial from the second sample chambers into the first sample chambers,performing the step of isolating the first sample chambers from thecorresponding second sample chambers.
 21. The method of claim 20,wherein the step of isolating the first sample chambers from thecorresponding second sample chambers is performed by physicallydeforming the radial fill conduits positioned between the first samplechambers and the second sample chambers.
 22. The method of claim 20,wherein the step of isolating the first sample chamber from thecorresponding second sample chamber is performed by one-way valvespositioned in the radial fill conduits.
 23. The method of claim 17,wherein the first biological material is a reagent and the secondbiological material is a sample to be tested.
 24. The method of claim17, wherein the first biological material is a sample to be tested andthe second biological material is a reagent.
 25. The method of claim 16,further comprising performing thermal cycling on the sample disk. 26.The method of claim 25, further comprising, simultaneously with the stepof performing thermal cycling, detecting the optical characteristics ofthe sample chambers.
 27. The method of claim 25, wherein the thermalcycling is performed without removing the sample disk from its positionon the rotational axis.
 28. The method of claim 25, wherein the step ofthermal cycling includes controlling the temperature of a sample blockthermally connected to the sample disk by a temperature control unitoperatively connected to the sample block.
 29. An apparatus forcentrifugally loading and thermally cycling a sample disk, comprising: asample disk having a plurality of first sample chambers, a plurality ofsecond sample chambers, and a reservoir for storing a volume of liquidsample; means for centrifugally loading the plurality of first samplechamber with liquid sample upon rotation of the sample disk about arotational axis of the sample disk; and means for centrifugally loadingthe plurality of first sample chambers with a biological material fromthe plurality of second sample chambers; and means for thermally cyclingthe plurality of first sample chambers of the sample disk.
 30. Theapparatus of claim 29, wherein the means for thermally cycling comprisesa sample block thermally connected to the sample disk.
 31. The apparatusof claim 30, wherein the means for thermally cycling further comprises atemperature control unit operatively connected to the sample block forraising and lowering the temperature of the sample block according to auser-defined profile.
 32. An apparatus configured for containing samplesof biological material during a thermal cycling operation, comprising: amicrocard configured for rotation about a rotational axis; a pluralityof first sample chambers positioned in the microcard around therotational axis; a plurality of second sample chambers positioned in themicrocard around the rotational axis, the second sample chambers beingpositioned closer to the rotational axis than the first sample chambers;and a plurality of channels formed in the microcard, the plurality ofchannels comprising a plurality of circumferential channels and aplurality of radial channels, the circumferential channels beingpositioned between adjacent first sample chambers to transport a firstbiological material from a reservoir into the plurality of first samplechambers upon rotation of the microcard about the rotational axis, theplurality of radial channels being positioned between correspondingfirst and second sample chambers to transport a second biologicalmaterial from the second sample chambers to the first sample chambersupon a further rotation of the microcard about the rotational axis,wherein the plurality of first sample chambers are configured to permitoptical detection of the biological materials in the first samplechambers.
 33. The apparatus of 32, wherein the plurality of first samplechambers are configured to permit thermal cycling during a rotation ofthe microcard about the rotational axis.
 34. The apparatus of claim 32,further comprising means for selectively blocking the passage of thefirst biological material through the circumferential channels after thefirst biological material has been transported into the first samplechambers.
 35. The apparatus of claim 34, the means for blocking thepassage of the first biological material comprising an elasticallydeformable region configured such that upon pressing a staking deviceagainst a surface of the microcard in the elastically deformable regionthe circumferential channels between adjacent first sample chambers isclosed off.
 36. The apparatus of claim 34, further comprising means forselectively blocking the passage of the first and second biologicalmaterials from the first sample chambers to the second sample chambers.37. The apparatus of claim 36, the means for selectively blocking thefluid communication of the first and second biological materialscomprising an elastically deformable region configured such that uponpressing a staking device against a surface of the microcard in theelastically deformable region the radial fill conduits between the firstsample chambers and the second sample chambers are closed off.
 38. Theapparatus of claim 32, wherein the microcard comprises a substantiallyflat circular plate.
 39. The apparatus of claim 32, wherein themicrocard is thermally connected to a sample block of a thermal cyclingdevice.
 40. The apparatus of claim 39, wherein the sample block isoperatively connected to a temperature control unit for raising andlowering the temperature of the sample block according to a user-definedprofile.